Nguyen Thu Giang vd Dig Tap chi KHOA HOC & CONG NGHE

Nguyen Thu Giang vd Dig Tap chi KHOA HOC & CONG NGHE 134(04): 6 9 - 7 3

NHAN GIONG CAY SAM 1>X\(Codonopsis javanica fBlume) Hook.f.) BANG KY T H U A T N U O I CAY M O THU'C V A T

NguySn Thu Giang , Tong Xuan Hoa ' Truang Dat hoc Y Duac - DH Thdi Nguyen.

'Tru&ng Dai hoc Su Pham - DH Thai Nguyen TOM T A T

Sam ddy la loai dugc lieu quy c6 rat nhieu cong dung chiJa benh vd phong ngira bfnh, giiip cho co the khde manh va sire dS khdng tot. Hifn nay, san lugng Sdm day khong dap ung dugc du nhu cdu trong nude. Vi vdy, viec nghien ciru quy trinh nhan nhanh cdy Sam ddy tao ra sd lugng Idn cay gidng sgch bfnh Id vifc lam rat can thidt. Sd dung phuang phap nuoi cdy mo td bdo thuc vdt, chiing t6i nhan thdy: Nhan nhanh choi cdy Sdm ddy trong moi truong MS co bd sung sucrose 25 g/l; agar 8,5 g/l; nude dua 50 ml/I; BAP 0,6 mg/l; 0,3 mg/l kinetin. Ty If bat chdi dat 4,66 chdi/mau, sau 4 tudn va 5,33 mdu/choi sau 8 tudn. Tao re cay tren MS co ban, c6 bd sung IBA 0,6 mg/I cho ty le chdi ra re dat 86,66% sau 8 tudn. Gia the ddt thjt trung binh co bd sung cdt, xa dira dd xu ly, phan chudng hoai theo ti If 2:1:2:1 va 5% NPK, 2% v6i bot co thd uom sdng 91% cay /n v//ro trudc khi chuyin cay ra moi truong ty nhi6n.

Tii khda' Sdm ddy, vi nhdn gidng, nuoi cdy md, BAP. Kinetin MCJDAU

Sam day hay cdn ggi la Dang sam ta logi dugc iifu quy duoc dua vao sach do Vift Nam td ndm 1996 [3]. Sam day ehua rat nhieu boat chat sinh hoc nhu: terpen, amin acid, hgp chat glycosid, vitamin, cae nguyen to khoang, alkaloid va hgp chat saponin...[1].

Sam day Id mgt vj thudc cd cdng dyng chira bfnh va phong ngda benh, giiip cho co the khoe manh va ed sue dk khang tdt. Cu sam day cd tac dung Igi tieu, nhuan trang, lam long ddm, than va re giup thanh nhift (ba sdt), lgi nieu, giai dpe[3]. Ddng y coi Sam day la nban sam ciia ngudi ngheo vi ed mpi cdng dyng cua nhan sam va co gia thdnh re [4] . Cung giong nhu nhieu loai eay thude quy khac, nan tan phd rimg Iam nuong ray qua mue trong ty nhien lam eho vimg pban bo cua cay Sam day bj thu h?p nhanh chong, cay co nguy ca bi cgn kift.

Ky thuat nuoi cay md te bao thye vat la mgt trong nbdng ky thuat dng dyng hifu qua trong nhieu ITnh vyc, dgc bift la nhan nhanh va bdo tdn cae lodi cay thudc quy cd gia trj kinh te cao[2], [8]. Bai bao nay eong bd kft qud nhan gidng cay Sam day bdng ky thuat nudi cay in vitro.

Tel: 0986 060359, Email: giangsinhdhyl23@gmail a

VAT LIEU VA PHLTONG PHAP Sd dyng cac dogn than mang chdi nacb ciia cae eay Sam day da dugc khd trung sach bfnh do Trgi thyc nghiem sinb hge Co Nhue - Td Liem - Ha Npi eung cap.

Nhan gidng in vitro cay Sam day theo mo ta cua D6 Nang Vinh (2005) [7]. Cae giai dogn nghien euu gdm nhan nhanh cay; tao re cay vd ra cay ngoai vudm uam.

Nghien cdu dugc thye hifn tren moi trudng MS CO ban co bo sung sucrose 25 g/l + agar 8,5 g/l + nude dua 50 ml/I va chat kieh thieh sinb trudng co nong dp thay ddi. Danh gia kha ndng nhan choi, tao re sau 4 tuan va 8 tuan nuoi cay. Tbi nghifm dugc tifn hanh d ciing difu kifn nbu nhau ve nhiet dp 25 ± 2''C, chieu sang 10/24 gid. Cac thi nghifm bo tri ngau nhifn, lap lai 3 lan

Phuong phap xd li sd lieu: Su dyng toan thong ke, cae sd lifu duge xd li trfn may vi tinb bdng chuang trinh Excel [6] . KET QUA VA THAO LUAN

Ngbien curu moi trirdng nhan choi Sam day Anh huang cua BAP (ten kha ndng nhan chdi vd sir sinh truang cUa choi cay Sam day trong ong nghiem

Nhan nhanh chdi la giai dogn gia tang so lugng choi trong mdt thdi gian ngan. Day la 69

Nguyin Thu Giang vd Dtg Tgp chi KHOA HOC & CONG NGriti giai doan quan trpng quyet dinh so lugng cay

trong qua trinh nuoi cay in vitro. Hf so nhan chdi cang cao thi kba nang nhan gidng cang ldn. He sd nhan chdi ehju anh hudng Idn ciia ydu td tac dgng la cae chdt kich thieh sinb trudng[5]. Cbung toi sd dung moi trudng MS CO bd sung BAP tu 0,2 mg/l dfn 1,0 mg/l de nghien cdu kba nang nhan ehdi va sinb trudng eua choi cay Sam day. Ket qua trinh bay d bang 1.

(3 bang I cbo thay, sau 4 tuan nudi eay sd chdi duge bat len td 1,33 den 4,66 choi/mau.

Sau 6 tudn, sd choi dat 1,58 den 5,33

kha nang nban cbdi, chieu cao choi eao hon so vdi cong thuc ddi chdng (edng thdc 1) khong bd sung BAP.

Xet ddng thdi cac chi tieu so cbdi/mSu, chieu eao chdi va chdt lugng chdi tbi chi tieu sd chdi/mau dugc coi la uu tien trong giai doan nhan nhanh. Do dd co the kft luan: cdngthiic 4 (bo sung BAP 0,6 mg/l vao mdi trudng nen MS + dudng 25 g/l + agar 8,5 g/l + nude dira 50 ml/1) la thfch bgp nhat trong thi nghifm, cho sd choi/mau dat 4,66 (sau 4 tuan nuoi edy) va 5,33 (sau 8 tuan nudi cdy), sy sinh trudng cua chdi dam bao, chat lugng chdi tot.

choi/mau. Cdc cong thuc e6 bo sung BAP cho

Bdng I. Anh hudng cua BAP den khd ndng nhdn choi vd su sinh irudng ciia choi Sdm ddy (sau 4 tudn vd 8 ludn)

Cong thuc

Nong df

BAP sd chdi/mSu Chieu cao

chdi (cm) Chdt lugng chdi

0,2 0,4 0,6 0,8 1,0

1,33 ±0,60 3,50 ± 0,37 4,00 ± 0,82 4,66 ± 0,28 2,83 ± 0,40 2,50 ± 0,47

1,4! ± 1,71 ± 2,41 ± 2,35 ± 2,00 ± 1,73 ±

Choi Chdi Chdi Chdi Chdi Chdi

gay hodc dj dang. Id xanh nhat hodc xodn hoi gay, la xanh

map. Id xanh map, Id xanh hoi gdy. Id xanh hoi gdy. Id xanh Sau 8 tu^V

gay hodc dj dgng. Id xanh nhat hodc xodn boi gay, Id xanh

mdp. Id xanh mgp. Id xanh 1(DC)

2 3 4 5 6

0 0,2 0,4

1,58 ±0,14 4,33 ± 0,44 4,50 ± 0,37 5,33 ±0,41 3,16±0,I9 3,33 ± 0,31

1,95 ± 4,00±

4,55 ± 4,08 ± 3,83 ± 2,66 ±

0,13 0,16 0r36 0,11 0,21 0,13

Choi Chdi Chdi Chdi

Chdi hai gdy, la xanh Chdi gay, la xann

hodc di dang. Id xanh nhgt hodc xodn

BAP 0,2 mg/I BAP Q^g mg/l

Hinh I. Anh hirdng aia BAP den khd ndng nhdn chdi vd susinh irudng cua chdi Sdm ddy (sau 8 ludn)A Anh huang cua kinetin din khd ndng nhan chdi vd su sinh truang cUa chdi Sam ddy irons ong nghiem

Kinetin Id mgt hgp chat co kha nang kich thich sy phan ehia td bao mgnh me thudng duge sit dyng trong nuoi eay md te bao thyc vat de lam tang hifu qua nhan ehdi va sy sinh trudng ciia chdi. Chung tdi tham do ndng dp kinetin tu 0,1 mg/l ddn 0,5 mg/l cbo kha ndng nhan chdi v^ sinh trudng cua ehdt cay Sam day, kdt qua trinh bay d bdng 2 .

Nguyen Tbu Giang vd Dtg Tap chi KHOA HOC & CONG NGH$ 134(04): 6 9 - 7 3 Bang 2. Anh hudng c 1 Kinetin den khd ndng nhdn choi vd su sinh trudng ciia choi Sdm ddy

(sau 4 tudn vd 8 ludn) C6ng

thdc Nong d$

Kinetin So choi/mau Chieu cao chdi

(cm) Chdt lirgng chdi

1(DC) 2 3 4 5 6

0 0.1 0,2 0,3 0,4 0,5

1,33 ±0,60 3,16±0,16 3,83 ±0,15 4,50 ± 0,27 2,83 ±0,21 2,16±0,35

1,41 ±0,08 2,66 ± 0,35 2,83 ±0,41 3,58 ±0,19 2,16±0,16 1,90 ±0,21

Ch6i gSy hoac di dang, la xanh nhat hoac xoan Choi hoi gay, 1^ xanh

Choi map, la xanh Choi mSp, la xanh Choi hoi gay. Id xanh

Choi gay hoac di dang, la xanh nhat hoac xoan Sau 8 tuan

l(BC) 2 3 4 5 6

0 0,1 0,2 0,3 0,4 0,5

1,58±0,14 3,66 ± 0,60 4,16±0,19 4,e6± 0,21 2,83 ±0,15 1,58 ±0,14

1,95 ±0,13 4,25± 0,32 4,41 ±0,32 6,66 ± 0,57 3,83 ± 0,32 1,95 ±0,13

Choi gay hoac di dang, la xanh nhat hoac xoan Choi hoi gay, la xanh

Choi map, la xanh Choi map, la xanh Choi hai gay, la xanh

Ch6i gay hoac dj dang, la xanh nhat hocic xoSn

Kinetin 0,1 mg/l Kinetin 0,3 mg/l Kinetin 0,5 mg/l li\n\i 2. Anh hudng cua Kinetin din khd nang nhdn choi vd su sinh trudng ciia chdi Sdm ddy (sau 8 tudn) Sau 4 tuan va sau 8 tuan nuoi eay dd lifu tai

bang 2 cho tbay cac cong thdc c6 bo sung kinetin cho kha nang nhan choi cao hon so vdi cong thdc ddi ehdng (cong thdc 1) khong bd sung kinetin. Khi bo sung kinetin vdi nong dp 0,3 mg/l d cdng thue 4 eho so choi/mau cao nhat dgt 4,50 (sau 4 tuan nudi cay) va 4,66 (sau 8 tudn nudi cay), sai khac co y nghia so vdi ddi cbiing. Sd chdi/mau tang ddn td ndng dp 0,1; 0,2 den 0,3mg/l va giam ddn tu ndng dp 0,3; 0,4 ddn 0,5. Vf chieu cao eiia ehdi edng thdc 4 cd gia trj Idn nhdt dgt 3,58 cm (sau 4 tudn nudi edy) va 6,66 cm (sau 8 tuan nuoi cay).

Ket hgp eae chi tieu ve s6 chdi/mau, chidu cao choi va ehat lugng choi ed the di den ket luan: cdng thue 4 co bd sung 0,3 mg/l kinetin vao mdi trudng nfn MS + dudng 25 g/l + agar 8,5 g/l + nude dda 50ml/l la thich hgp nhdt trong thi nghifm, cho sd chdi/mau dat cao, chdt lugng chdi tdt.

Nghien cdu moi trirdng t^o re cua chdi Sam day

Sdm day cao khoang 4 - 5 cm dugc cay tren mdi trudng MS eg ban co bd sung 25g dudng, 8,5g thach aga va IBA (0,2mg/l; 0,4mg/l;

0,6mg/l; 0,8mg/l). Chung tdi da tien hdnh tham dd anb hudng cua IBA len sy phat sinh re ciia Sam day va thu duge ket qua theo bang so lieu dudi day. Ket qua d bang 3 eho thay, cac cong thuc thi nghiem co bd sung IBA vdi ndng do td 0,2mg/l - 0,4 mg/l - 0,6mg/t - 0,8mg/l deu eo sy ra re vdi cac muc dp phat trien khac nhau. O cdng tbdc doi chdng tbi khdng phat sinh re. Bd sung IBA 0,6 mg/l cho re dep, mau vang va so lugng re nhieu nhat dgt 7,30 re/cay sau 8 tuan nudi eay. Con d cac cong thdc moi trudng khac co phat sinh re nhung re binh thudng, mau trang hodc re xau.

Nhu vay, moi trudng thich hpp nhat cbo sy tao rf ciia chdi Sam day la MS + 25g dudng + 8,5aga+ lBA0,6mg/l.

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Nguyin Thu Giang vd Dtg Tgp chi KHOA HOC & CONG NGHE

Bang 3. Anh hudng cua IBA din khd ndng lao re ciia chdi Sdm ddy (sau 8 tudn) C6ng Nong df

thirc [BA (mg/l)

Ti ie chfii ra re

(%)

So re /cdy Chdt Iir9ng rl 1(DC)

2 3 4 5

0,0 0,2 0,4 0,6 0,8

0,00 ± 0,00 0,00 ± 0,00 Khong ra rl

68,33 ± 4,40 4,50 ± 0,51 Bd mJt rl tron nhdn, mdu trdng 76,66 ± 4,40 5,73 ± 0,89 B I mdt rl trrni nhdn, mdu vdng 86,66 ± 1,66 7^30 ± 0,36 BS mdt rl trcm nhdn, mdu vdng 71,66 ±3,33 4,70 ± 0,96 Bd mdt rl trcm nhdn, mau trdng

Khong bo sung IBA 86 sung IBA 0,4 mg/l Bd sung IBA 0,6 mg/l Hinh 3. Anh huang cua IBA den khd ndng tgo ri cua choi Sdm ddy (sau 8 ludn) Ket qua dira cay ra moi trirdng tur nhien

Sau 8 tudn tgo rf, khi cay Sam day cao khoang 4 - 5em va c6 khoang 7 - 8 rl chimg tdi tifn hanh dua cay ra moi trudng ty nhien. Chung toi sd dyng 3 gia the va tbeo doi cac cbi tieu sau 4 tuan ehdm soc (bang 4).

Bang 4. Anh hudng cua gid the den li le song vd su sinh truang cua cdy Sdm ddy in vitro Irong vudn uom (sau 4 ludn)

Cong

Ihuc Gid the Ti If cdy s6ng (%) Chieu cao cay (cm)

I (DC) Ddt thit trung binh 2 Ddt thjt trung binh + cdt (2:1)

Dat thjt trung binh + cdt + xa dua da xir ly + 3 phdnchudnghoai theoti le2:l:2:l bd sung 5%

NPKvd2%v6ibQt

60,00 ± 11,54

77,33 ± 1,45 11,36 ±0,79 12,99 ±0,51

Cdng thirc 3 fsau 4 ludn) Cdng thdc 1 Cong thirt 2

Hinh 4. Cdy Sdm ddy dua ra mui nuung u

Ket qua d bang 4 eho thay, ti If cay sdng d cac gia thd sau 4 tuan trdng dao dpng tu 60% dfn 91%. Tren gia the ddt thjt trung binh vdi ti If sdng dgt 60%, eay sinb trudng ydu ban so vdi cac eong thuc kbdc, chidu cao eay chi dat 11,36 ± 0,79 cm. Tren gia thd ddt thjt trung binh co bd sung cat, xo dua dd xu ly, pban chudng hoai theo ti If 2:1:2:1 vd 5 % NPK, 2% vdi bpt, ti If cay s6ng dgt 91%, cay map, khoe, la to, xanh tham, day ban, chidu cao cay dgt 18,06 ± 1,64 cm.

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Nguyen Thu Giang vd Dtg Tap chi KHOA HQC & C 6 N G NGHE 134(04): 69-73 Nhu vay, trong pham vi cua thi nghiem, gia

the phii bgp giai doan luyen cay trong vudn uam la dat thjt trung binh + eat + xa dua da xd ly + phan chuong hoai theo ti le 2:1:2:1 bd sung 5% NPK va 2% voi bgt

KET LUAN

Mdi tiudng thieh hgp nhdt eho nhan nhanh va sy sinb trudng ehdi Sam day la MS co bd sung sucrose 25 g/l; agar 8,5 g/l; nude dua 50 mi/1;

BAP 0,6 mg/l; 0,3 mg/l kinetin; pH = 5,8.

Moi trudng thicb hgp nhdt eho kha nang tgo re eay Sam day la MS cd bo sung sucrose 25 g/l; agar 8,5 g/l; nude dda 50 ml/I; IBA 0,6 mg/l; pH = 5,8.

Gia the thieh hgp nhat cho cay Sam day sinh trudng d giai dogn vudn uam trudc kbi trdng eay trong san xuat la dat thit trung binh eo bo sung edt, xa dda da xd ly, phan chudng hoai theo ti If 2:1:2:1 va 5% NPK, 2% voi bgt.

TAI LIEU THAM K H A O I. Hodng Minh Chung, Pham Xuan Sinh, Nguyen Manh Tuyen (2002), "Budc dau nghien cii'u thdnh

phdn boa hpc ciia vi thuoc Dang sam vift nam", Tgp chi duac li4u, 7 (l), tr: 36.

2. Nguyin Vift Cutmg, Ho Thanh TSm, Nguyin Bd Nam, Ha Thj My Ngan, Le Kim Cuong, Nguyen Phuc Huy, Duong Tdn Nhyt (2013),

"Nghi6n cdu anh huong mot so chat httu co va bac Nitrat len sy phdt trien cua cay sdm Ngpc Linh (Panax Viemamensis Ha Et Grushv.) nu6i cdy in vitro, Hgi nghi KHCN sinh hge loan qude. Ha Nfi 27/9/2013

3. Nguyen Thugng Dong, Tran Cong Lugn, Nguyin Thj Thu Huang (2007), Sdm Vi^t Nam vd mgl so cdy thudc ho Nhdn sdm, Nxb Khoa hge vd Ky thugt.

4. Le Trdn Ddc, (1983), Trdng hdi vd diing cdy thuoc, Tap 1, Nxb N6ng Nghifp

5. Ngo Thanh Tdi, Nguyin Bd Nam, H6 Thanh Tdm, Hd Thj My Ngdn, Duong Tdn Nhut (2013),

"Nghien ciiu tdc dong cua dnh sdng d6n LED Ien khd ndng tdng sinh mo s^o va sy hinh thanh cay hodn chinh tu phoi vo tinh cay sdm Ngpc Linh (Panax vietnamensis Ha ET Grushv.)", Bdo cdo khoa hge - Hgi nghi khoa hge cdng nghf sinh hge todn quoc, Nxb Khoa hge ty nhi^n vd c5ng nghf, U. 1038-1042.

6. Nguyin Hai Tudt, Ngo Kim Kh6i i\996),Xuli ihdng ke kel qud nghien cuu thuc nghiem Irong ndng ldm ngu nghiep tren mdy vi tinh, Nxb Nong nghifp, HaN6i.

7. Dd Ndng Vinh (2005), Cong nghe ti bdo thuc vdt ling dung. Nxb Nong nghifp.

SUMMARY

USING IN VITRO PLAN TISSUE CULTURE METHOD TO MULIPLY CODONOPSIS JA VANICA

Nguyen Thu Giang'*, Tong Xuan Hoa ' 'College of Medicine and Pharmacy - TNU, ^College cf Education - TNU Codonopsis javanica are medicines that have a lot of healing and diseases prevention, help the body to be healthy and have good resistance. Currently, Sam wire output are not enough to meet domestic demand. Therefore, the research in Sam's fast wireless plant process generate large numbers of fresh seedling is very necessary work. Using plant cells transplant method we found that: Sam's fast wireless shoots in MS medium supplemented with sucrose 25 g/l; agar 8,5 g/l;

Coconut water 50 ml/I; BAP 0.6 mg/l; 0.3 mg/I kinetin. The rate reached 4.66 out bud/form, after 4 weeks and 5.33 shoots /sample after 8 weeks. Create roots in basic MS, supplemented with IBA 0.6 mg/l ratio reached 86.66% of rooted shoots after 8 weeks. Prices may average soil - sand - coconut coir processing - manure at the rate of 2: 1:2: 1 and 5% NPK, 2% lime can live 91% tree nursery in vitro before moving trees to the natural environment.

Key words: Codonopsis javanica, in vitro cultured, BAP, kinetin.

Ngdy nhgn bdi. 08/12/2014: Ngdy phdn biin:22/12/2014; Ngdy duyet ddng: 08/5/2015 Phdn bien khoa hoc: TS. Nguyen Thu Hiin - Trudng Dgi hge Y Dugc - DHTN

Tei 0986 060359, Email giangstnhdhyl23@gmail.c>